Ma Lieting, Liu Wenyan, Li Miaoling
Department of Laboratory Medicine, The First Affiliated Hospital of Xi'An Jiaotong University, Xi'an, 710061, Shaanxi, China.
Department of Hemodialysis, The First Affiliated Hospital of Xi'An Jiaotong University, Xi'an, 710061, Shaanxi, China.
Biochem Genet. 2023 Apr;61(2):628-650. doi: 10.1007/s10528-022-10277-6. Epub 2022 Sep 2.
Previous studies have suggested that circular RNAs (circRNAs) play important regulatory roles in cancer progression. Previous evidence exhibited the aberrant upregulation of circ_0061140 in ovarian cancer. However, the detailed role of circ_0061140 in ovarian cancer progression and its associated mechanism remain largely unknown and need further exploration. The expression of circ_0061140, microRNA-761 (miR-761) and leucine zipper and EF-hand containing transmembrane protein 1 (LETM1) was checked by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) or western blot. Cell Counting Kit-8 (CCK8), colony formation, 5-Ethynyl-2'-deoxyuridine (EdU), flow cytometry, wound healing, transwell, and tube formation assays were conducted to assess cell functions. Dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay were performed to confirm the interaction between miR-761 and circ_0061140 or LETM1. Xenograft tumor model was established to analyze the role of circ_0061140 in tumor growth in vivo. Circ_0061140 expression was notably up-regulated in ovarian cancer tissues and cell lines. Circ_0061140 knockdown suppressed the proliferation, migration, invasion, and angiogenesis and triggered the apoptosis of ovarian cancer cells. Circ_0061140 directly interacted with miR-761, and circ_0061140 silencing-mediated anti-tumor effects were partly abolished by miR-761 knockdown in ovarian cancer cells. LETM1 was a direct target of miR-761, and LETM1 overexpression partly counteracted miR-761-induced anti-tumor effects. Circ_0061140 could up-regulate LETM1 expression by sponging miR-761. Circ_0061140 knockdown significantly suppressed xenograft tumor growth in vivo. Circ_0061140 aggravated ovarian cancer progression through miR-761-dependent regulation of LETM1.
先前的研究表明,环状RNA(circRNAs)在癌症进展中发挥重要的调控作用。先前的证据显示,circ_0061140在卵巢癌中异常上调。然而,circ_0061140在卵巢癌进展中的具体作用及其相关机制在很大程度上仍不清楚,需要进一步探索。通过逆转录-定量聚合酶链反应(RT-qPCR)或蛋白质免疫印迹法检测circ_0061140、微小RNA-761(miR-761)和含亮氨酸拉链和EF手结构域的跨膜蛋白1(LETM1)的表达。进行细胞计数试剂盒-8(CCK8)、集落形成、5-乙炔基-2'-脱氧尿苷(EdU)、流式细胞术、伤口愈合、Transwell和管形成试验以评估细胞功能。进行双荧光素酶报告基因测定和RNA免疫沉淀(RIP)测定以证实miR-761与circ_0061140或LETM1之间的相互作用。建立异种移植肿瘤模型以分析circ_0061140在体内肿瘤生长中的作用。circ_0061140在卵巢癌组织和细胞系中明显上调。敲低circ_0061140可抑制卵巢癌细胞的增殖、迁移、侵袭和血管生成,并引发细胞凋亡。circ_0061140直接与miR-761相互作用,在卵巢癌细胞中,敲低circ_0061140介导的抗肿瘤作用部分被敲低miR-761所消除。LETM1是miR-761的直接靶点,LETM1过表达部分抵消了miR-761诱导的抗肿瘤作用。circ_0061140可通过海绵吸附miR-761上调LETM1表达。敲低circ_0061140可显著抑制体内异种移植肿瘤的生长。circ_0061140通过miR-761依赖性调控LETM1加重卵巢癌进展。